Device for measurement of transport systems

Abstract

The invention relates to a device ( 1 ) for optically measuring properties of transport systems ( 50 ) in membranes ( 40 ), particularly carrier or channel proteins. In order to be able to measure the properties of biological transporter molecules ( 50 ) with a high throughput, the invention proposes that the device ( 1 ) comprise an optical measurement unit ( 2 ) and a data processing unit ( 6 ) having a process control ( 7 ) and a data capture.

1 . Device ( 1 ) for optical measurement of properties of transport systems in membranes, particularly of carrier proteins or channel proteins, as well as secretion mechanisms, wherein it has an optical measurement device ( 2 ), a data processing unit ( 6 ) for capture and evaluation of measurement data, and a process controller ( 7 ). 
     
     
         2 . Device ( 1 ) according to  claim 1 , wherein the optical measurement device ( 2 ) is a TIRF microscope. 
     
     
         3 . Device ( 1 ) according to  claim 1 , wherein a sample manipulator ( 4 ) that can be controlled by the process controller ( 7 ) is provided for the device ( 1 ). 
     
     
         4 . Device ( 1 ) according to  claim 1 , wherein the device ( 1 ) is set up for measuring biochips ( 9 ) that have a transparent layer ( 20 ) having multiple measurement chambers ( 30 ). 
     
     
         5 . Device ( 1 ) according to  claim 1 , wherein the optical measurement device ( 2 ) has a beam guide whose angle is smaller than a TIRF angle, preferably at most smaller than 20% of the TIRF angle. 
     
     
         6 . Device ( 1 ) according to  claim 1 , wherein the device ( 1 ) has an incubation station ( 8 ) for the biochips ( 9 ) to be measured, which can be controlled by the processor controller ( 7 ). 
     
     
         7 . Device ( 1 ) according to  claim 1 , wherein the device ( 1 ) is configured to carry out measurement cycles with process control and in automated manner, whereby a measurement cycle, in each instance, essentially comprises prepping of the biochip ( 9 ) for the measurement, a subsequent optical measurement, and a subsequent evaluation of the measurement data. 
     
     
         8 . Device ( 1 ) according to  claim 1 , wherein the sample manipulator ( 4 ) is configured for carrying out the following steps on the biochip ( 9 ):
 a) equilibration with a buffer solution,   b) addition of proteo-liposomes, biological membranes, or cells,   c) washing with buffer solution,   d) addition of substrates and/or active substance candidates,   e) placement in the measurement region.   
     
     
         9 . Device ( 1 ) according to  claim 1 , wherein the device ( 1 ) is configured for carrying out a time-resolved fluorescence measurement of the measurement chambers ( 30 ) of the biochip ( 9 ) after prepping of the biochip ( 9 ), and for capturing and processing the measurement data. 
     
     
         10 . Device ( 1 ) according to  claim 1 , wherein the data processing unit ( 6 ) is set up for carrying out the following steps after the fluorescence measurement:
 a) determination of the time-resolved fluorescence intensity for the individual measurement chambers ( 30 ), in each instance, by means of pattern recognition,   b) fitting of a mathematical curve to the time-resolved fluorescence intensity,   c) classification of the measurements chambers ( 30 ), on the basis of the mathematical curve, in one of the three following categories: i) sealed measurement chambers with a fluorescence signal, ii) sealed measurement chambers without a fluorescence signal, iii) open measurement chambers, and   d) rejection of the measurement data of sealed measurement chambers without a fluorescence signal and of open measurement chambers, and   e) calculation of a parameter for the velocity of transport, preferably of the velocity constant for transport, for each sealed measurement chamber with a fluorescence signal.   
     
     
         11 . Device ( 1 ) according to  claim 1 , wherein the data processing unit ( 6 ) is set up for performing the following steps:
 a) plotting of all calculated velocity constants for transport against their frequency in a histogram,   b) assignment of a corresponding number of transport systems per measurement chamber ( 30 ) to the velocity constants, in each instance,   c) determination of a specific velocity constant of a transport substrate ( 60 ) used for a transport system ( 50 ) per measurement chamber ( 30 ).   
     
     
         12 . Method for optical measurement of properties of transport systems ( 50 ) in membranes ( 40 ), particularly of carrier proteins or channel proteins, as well as secretion mechanisms, which comprises the following steps:
 a) determination of the time-resolved fluorescence intensity for individual measurement chambers of a biochip ( 9 ), in each instance, by means of pattern recognition,   b) fitting of a mathematical curve to the time-resolved fluorescence intensity,   c) classification of the measurement chambers ( 30 ), on the basis of the mathematical curve, in one of the three following categories: i) sealed measurement chambers with a fluorescence signal, ii) sealed measurement chambers without a fluorescence signal, iii) open measurement chambers.   
     
     
         13 . Method according to  claim 12 , wherein it comprises the following steps:
 d) rejection of the measurement data of sealed measurement chambers without a fluorescence signal and of open measurement chambers, and   e) calculation of a parameter for the velocity of transport, preferably of the velocity constant for transport, for each sealed measurement chamber with a fluorescence signal.   
     
     
         14 . Method according to  claim 12  wherein it comprises the following steps:
 f) plotting of all calculated velocity constants for transport against their frequency in a histogram,   g) assignment of a corresponding number of transport systems ( 50 ) per measurement chamber ( 30 ) to the velocity constants, in each instance,   h) determination of a specific velocity constant of a transport substrate used for a transport system ( 50 ) per measurement chamber ( 30 ).